CN114410765A - Application of miRNA molecular marker in schizophrenia detection - Google Patents

Application of miRNA molecular marker in schizophrenia detection Download PDF

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CN114410765A
CN114410765A CN202110555900.2A CN202110555900A CN114410765A CN 114410765 A CN114410765 A CN 114410765A CN 202110555900 A CN202110555900 A CN 202110555900A CN 114410765 A CN114410765 A CN 114410765A
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赵春华
王娇
黄琳
史宏伟
齐雯歆
刘翠萍
张诗博
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University of Shanghai for Science and Technology
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Abstract

The invention discloses an application of miRNA molecular marker in schizophrenia detection, which analyzes the whole genome difference of gene expression between a schizophrenia patient and a healthy control group by using a bioinformatics method, and then analyzes the signal path and function of the difference gene participation by GO and KEGG functional enrichment; then establishing a cerRNA network to analyze the interaction between the upstream key miRNA and the upstream key miRNA including the central target gene RASD2 and predict the optimal binding site of the upstream key miRNA and the central target gene RASD 2; the expression of DEGs is regulated by miRNA, and hsa-miRNA-4763-3p targeting RASD2 gene is used as a potential biomarker and a therapeutic target of schizophrenia. The miR-4763-3p can be used as a potential biomarker and a therapeutic target of schizophrenia, and provides a theoretical basis for further clinical study of schizophrenia.

Description

Application of miRNA molecular marker in schizophrenia detection
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a schizophrenia molecular marker miRNA-4763-3p and application thereof in the fields of screening of high risk groups of schizophrenia, early diagnosis and treatment of schizophrenia and the like.
Background
Schizophrenia is a common serious mental disease with disorder in perception, thinking, emotion, behavior and the like and uncoordinated mental activities, has more than 2000 million people in global influence, has high morbidity in adolescence or early adulthood, and has the early mortality rate 2-3 times higher than that of common people. The typical clinical symptoms are hallucinations and delusions, patients are isolated in early onset and do not want to communicate with others, the disease condition is further developed, namely thought disorder and fantasy auditory hallucinations appear, and some patients can also have suicide, self-disabled people and impulsion of leaving home. The disease seriously damages the mental and physical health of patients and brings heavy burden to families and society of the patients. Therefore, it is necessary to find a technical means for preventing and treating schizophrenia.
MicroRNAs (miRNAs) are small non-coding RNA molecules, are degraded or mediate the translational inhibition of target mRNA by complementary or partially complementary combination with a 3 'untranslated region (3' UTR), and participate in various biological processes such as cell proliferation, apoptosis, differentiation, metabolism, development, tumor metastasis and the like. mirnas play a crucial role in brain development and the pathophysiology of many psychiatric diseases. Studies of miRNA expression in the prefrontal cortex at different physiological stages in schizophrenic patients reveal the spatiotemporal dynamics of mirnas in this region, suggesting that there may be a link between schizophrenia and mirnas in infant and adolescent dysfunction. miRNAs and derivatives thereof have great prospects in disease diagnosis and treatment.
Biomarkers are biochemical markers that can label pathological changes of cells, tissues, organs at the molecular level. Due to the advantages of sensitivity and specificity, the kit can provide early warning and prognostic efficacy analysis and can also be used as a basis for auxiliary diagnosis to a great extent. A number of techniques have been used to find biomarkers for schizophrenia. Recent studies have shown that mirnas may be attractive candidate biomarkers as modulators of gene expression due to their stability and specificity. In particular, mirnas in whole blood or specific blood components are candidate drugs for improving disease diagnosis, including life-threatening pathologies. The core role of blood mirnas in the pathogenesis of schizophrenia has also been found in studies related to schizophrenia. However, the application of miRNA molecules in biomedical detection of schizophrenia is still in the first stage, and new and more effective solutions are needed for their application to fully function.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to overcome the defects in the prior art, and provides an application of a miRNA molecular marker in schizophrenia detection, wherein the whole genome difference of gene expression between a schizophrenia patient and a healthy control group is systematically analyzed by using a bioinformatics method, and then the signal path and function in which the different genes participate are analyzed by GO and KEGG function enrichment. Next, a cerana network was constructed to analyze the interaction between upstream critical mirnas and their inclusion of the central target gene RASD2, their optimal binding sites predicted using RNAhybrid and BLASTN software. Finally, the molecular mechanism of schizophrenia is analyzed and verified in a mouse model and schizophrenic patients through molecular experiments.
In order to achieve the purpose, the invention adopts the following inventive concept:
the present invention uses bioinformatic methods to systematically analyze genome-wide differences in gene expression between schizophrenic patients and healthy controls, as well as the signaling pathways and functions in which these differential genes participate. Next, a ceRNA network was constructed to analyze the interaction between the upstream key miRNA and its central target gene RASD2 and predict their optimal binding sites. In addition, mirnas were found to regulate the expression of these DEGs, targeting hsa-miRNA-4763-3p of RASD2 gene as potential biomarker and therapeutic target for schizophrenia. These results were finally validated in mouse models and patient blood samples. The research is based on bioinformatics analysis of DEGs and validation of blood samples, and tries to discover miRNAs as potential biomarkers of schizophrenia, thereby laying a clinical foundation for early diagnosis and treatment of schizophrenia.
According to the inventive concept, the invention adopts the following technical scheme:
the application of miRNA molecular marker in schizophrenia detection is characterized in that the whole genome difference of gene expression between a schizophrenia patient and a healthy control group is analyzed by using a bioinformatics method, and then the signal path and function in which the different genes participate are analyzed by GO and KEGG functional enrichment; then establishing a cerRNA network to analyze the interaction between the upstream key miRNA and the target genes thereof, including the central target gene RASD2 and predict the optimal binding sites of the upstream key miRNA and the target genes; the expression of DEGs is regulated by miRNA, and hsa-miRNA-4763-3p targeting RASD2 gene is used as a potential biomarker and a therapeutic target of schizophrenia. The molecular mechanism of schizophrenia is analyzed and verified in a mouse model and schizophrenic patients through molecular experiments, and the molecular mechanism of schizophrenia is discussed at a molecular level, so that people can find molecular markers for early diagnosis of schizophrenia, and a new target is provided for early prevention and treatment of schizophrenia.
In order to determine the gene difference between the schizophrenic patients and the healthy control group, the RNA-Seq sequencing results of human ipsc-derived cortical interneurons in the two groups were analyzed, and 61 DEGs were determined in 28 schizophrenic patients and 28 healthy control groups. Of these 61 DEGs, 21 genes were up-regulated in schizophrenia patients, such as C1orf54 and HLA-B. There are 40 genes down-regulated, such as SETD7, MYO5B, KCTD12 and RASD 2.
Preferably, the functional enrichment analysis is performed by the gene ontology GO (https:// david. ncifcrf. gov /) and the gene and genome encyclopedia KEGG, the GO analysis comprising:
(1)17 genes were enriched in the biological process: including protein homooligomerization, modulation of potassium ion transport across membranes, axonal guidance, modulation of protein kinase activity, and retinal homeostasis;
(2)29 genes were enriched in cellular components: including cell junctions, cell surfaces, extracellular vesicles and golgi apparatus;
(3)13 genes are enriched in molecular functions: comprises voltage-gated potassium channel activity, polysaccharide binding, RNA polymerase II sequence specific DNA binding and potassium channel activity;
the KEGG analysis involved 10 DEGs participating in five important pathways: graft versus host disease, allograft rejection, aldosterone-mediated sodium resorption, nicotine addiction, and type I diabetes.
Preferably, the GO analysis of the present invention shows: 17 genes are enriched in a biological process, 29 genes are enriched in cell components, 13 genes are enriched in molecular functions, and KEGG analysis shows that 10 DEGs participate in five important ways to obtain the relationship between the DEGs and schizophrenia.
Preferably, the intermediate node of the cerRNA network, miRNA, regulates the expression of DEGs and related lncRNA, and the upstream miRNA of RASD2 gene comprises has-miRNA-4763-3p, hsa-miR-6820-5p, hsa-miR-6756-5p, hsa-miRNA-4695-5p and hss-miRNA-6769b-5p, which form the backbone of the whole network.
The invention utilizes the molecular mechanism of DEGs related to schizophrenia, and the intermediate node miRNA of the cerRNA network regulates the expression of the DEGs and related lncRNA. This result suggests that aberrant expression of DEGs in schizophrenia may be regulated by various mirnas. In addition, the most upstream miRNA of RASD2 gene forms the central core of the entire network, and these mirnas are involved in regulating the expression of RASD2 in schizophrenia.
Preferably, images are constructed and binding sites of target genes are predicted using RNAhybrid and BLASTN.
Preferably, hsa-miRNA-4763-3p is used as the optimal target site for binding to RASD 2.
The binding free energy between hsa-miRNA-4763-3p and RASD2 adopted by the invention is the lowest, and the result shows that the interaction exists between miR-4763-3p and RASD2, and hsa-miRNA-4763-3p is the optimal target site for RASD2 binding.
Preferably, key brain areas are analyzed for specific detection of schizophrenia by injecting 5-HT antagonist 18F-MPPF for specific detection of expression profile of 5-HT receptors in schizophrenic patients, and key brain area information is obtained.
In combination with the PET data of schizophrenic patients, 5-HT can modulate the release of certain neurotransmitters, including dopamine, in the prefrontal cortex. The injection of the highly selective 5-HT antagonist 18F-MPPF can be used to specifically detect the expression profile of 5-HT receptors in schizophrenic patients. PET data were collected from 44 specimens (including 18 schizophrenic patients and 26 healthy controls) and analyzed using MATLAB R2016 a. By comparing data from schizophrenic patients and healthy individuals, we obtained key brain regions. The increased brain areas are marked red and the decreased brain areas are marked blue, which means that the expression of 5-HT receptors in the brain of schizophrenic patients is up-regulated mainly in the frontal, temporal and down-regulated in the occipital, posterior and left temporal lobes.
Preferably, hsa-miRNA-4763-3p is used as a candidate biomarker for schizophrenia.
The invention researches the important function of RASD2 in schizophrenia and verifies the relation between miR-4763-3p and RASD 2. qPCR analysis was first performed in the mouse schizophrenia model, confirming the differential expression of RASD 2. Subsequently, qPCR analysis of blood samples from schizophrenic patients showed that the expression level of RASD2 was down-regulated, hsa-miRNA-4763-3p and hsa-miRNA-6769b-5p targeted to RASD2 was up-regulated, and hsa-miRNA-6820-5p was down-regulated. Our findings suggest that these miRNAs may be candidate biomarkers for schizophrenia.
Compared with the prior art, the invention has the following obvious and prominent substantive characteristics and remarkable advantages:
1. the molecular mechanism of schizophrenia is discussed at the molecular level, so that people can find molecular markers for early diagnosis of schizophrenia, and a new molecular target is provided for early prevention and treatment of schizophrenia; in the present invention, a bioinformatics method was first used to systematically analyze the whole genome differences in gene expression between schizophrenic patients and healthy controls, and 61 Differentially Expressed Genes (DEGs) were initially screened in schizophrenic patients; GO and KEGG functional enrichment analysis shows that most genes are mainly enriched in related functions of synaptic transmission and neurotransmitter transmission; then, a ceRNA network is constructed to analyze the interaction between the upstream key miRNA and the target gene thereof, wherein the research finds that the number of the upstream miRNA of the RASD2 gene is the maximum, and the central nervous system of the whole network is formed; the results show that different miRNAs regulate the expression of the DEGs, so that miR-4763-3p can regulate the expression of a target gene RASD2 of the miRNAs, thereby influencing the occurrence and development of schizophrenia; next, the results of PET show the key brain regions of schizophrenia pathology such as prefrontal lobe, etc. and qPCR analysis proves the differential expression of RASD2 in mouse models of schizophrenia, thus proving the key regulation and control function of RASD2 in schizophrenia; finally, qPCR analysis is carried out on blood samples of schizophrenic patients, and expression of RASD2 is reduced, and expression of miR-4763-3p expressed by target RASD2 is increased;
2. the miR-4763-3p can be used as a potential biomarker and a therapeutic target of schizophrenia, and provides a theoretical basis for further clinical study of schizophrenia.
Drawings
FIG. 1 is a screening chart of the differential genes associated with schizophrenia. Wherein, figure (a): gene expression difference analysis heatmap of schizophrenic patients (n-28) and healthy control group (n-28), red indicates up-regulated genes and blue indicates down-regulated genes. The DEG screening threshold is: | log2FC | >0.38, p value < 0.05. (RNA sequencing data downloaded from GEO (https:// www.ncbi.nlm.nih.gov/GEO/; accession number GSE 121376.) figure (b): differential Gene volcano graph.Red represents up-regulated genes and Green represents down-regulated genes.Total 61 differential genes were present, 21 of which were up-regulated and 40 of which were down-regulated.figure (c): the up-and down-regulation of the expression of differential genes that have been shown to be associated with schizophrenia.
Previously reported RNA-Seq data for schizophrenic patients and healthy individuals were downloaded from the GEO database, setting | log2FC | >0.38, with a p-value <0.05 as the threshold for screening for differential genes.
FIG. 2 is a functional enrichment analysis diagram of the differential gene GO/KEGG. Wherein, FIGS (a-c): functional annotation of differential genes in biological pathways, cellular components, and molecular functions. 17 genes participate in biological processes such as protein homologous oligomerization, potassium ion transmembrane transport regulation, axon guidance, protein kinase activity regulation, intraretinal homeostasis and the like; 29 genes are enriched in cell components such as cell surfaces, Golgi bodies and the like; the 13 genes perform molecular functions such as voltage-gated potassium channel activity, polysaccharide binding, RNA polymerase II sequence-specific DNA binding, and the like. FIG. (d): the differential gene KEGG pathway analysis mainly relates to 5 important pathways: graft versus host disease, allograft rejection, aldosterone-mediated sodium reabsorption, nicotine addiction, and type I diabetes. FIG. (e): key genes are in the major pathways and functions involved and performed.
The selected gene differences related to schizophrenia were annotated for GO/KEGG function using DAVID 6.8(https:// DAVID. ncifcrf. gov /).
FIG. 3 is a diagram of the analysis of the ceRNA network of the differential genes. The RNAhybrid and BLASTN were used to determine the regulatory relationship between miRNA-mRNA and miRNA-lncRNA. The light blue represents the node has-miRNA, the red node represents up-regulated mRNA and the dark blue node represents down-regulated mRNA. The orange node represents up-regulated lncRNA, and the green node represents down-regulated lncRNA. The gray lines represent the interaction between the two nodes. With RASD2 in an intermediate position in the network.
FIG. 4 is a graph showing the prediction of the relationship between miRNA and mRNA. Wherein figure (a): the values for at least four different miRNA-regulated protein-encoding genes are shown for the targeted prediction of four different miRNA-schizophrenia related genes. FIG. (b): binding sequences of miRNA and RASD2 were predicted by RNAhybrid software. FIG. (c): RPKM values for six genes in schizophrenic and healthy controls.
Figure 5 is a PET image of the brain of a schizophrenic patient. Use of schizophrenia markers18PET imaging of 5-HT receptors by F-MPPF, red for enhanced brain regions and blue for diminished brain regions.
Fig. 6 is a qPCR validation expression profile of key genes and mirnas. Wherein figure (a): of the mouse brain18F-DTBZ PET images; green represents dopamine receptor. FIG. (b): qPCR detection of gene expression in mice of the schizophrenic model and in healthy control mice RASD2, KCTD12 and SETD 7. FIG. (c): qPCR detectionRASD2 gene expression in blood samples of schizophrenic patients and healthy subjects. FIG. (d): qPCR detects the expression of mirnas involved in regulating RASD2 gene in blood samples of schizophrenic and healthy subjects. Data expressed as mean ± SEM, compared to control group,. p<0.05；**P＜0.01；***P＜0.001。
A schizophrenic mouse model was constructed by intraperitoneal injection of the N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 into mice for one week. After the neck is cut off and the mouse is killed, the prefrontal lobe of the mouse is quickly taken, RNA is extracted by using a total RNA extraction kit, and then the RNA is reversely transcribed into cDNA. qPCR was performed to quantify the expression level of genes involved in schizophrenia.
Blood samples were collected using a blood collection tube without anticoagulant, and serum was taken after blood coagulation, after RNA was extracted using a total RNA extraction kit, and cDNA was reverse transcribed for detection as above.
Detailed Description
The above-described scheme is further illustrated below with reference to specific embodiments, which are detailed below:
the first embodiment is as follows: acquisition and functional annotation of differentially expressed genes:
to identify differentially expressed genes between schizophrenic and healthy individuals, previously reported RNA-Seq data (GEO accession No. GSE121376) were obtained. Low quality bases at the 5 'and 3' ends (Q20 ≦ 20) were deleted using locally developed Perl script. The read fragments were aligned to the human reference genome (grcm38.p5) using HISAT2 (version 2.0.5). Reads were performed and counted using BEDTools (Quinlan 2014), and their expression was calculated using RPKM (reads per million bases) method. Differentially expressed genes were screened using a fold change threshold of | log2FC | >0.4 and p-values were obtained by t-test. Subsequently, DAVID 6.8 is used for carrying out function enrichment analysis on the schizophrenia-related gene.
Example two: construction of the cerRNA network:
collecting miRNA from miRbase. The mRNA and lncRNA sequences were obtained from NCBI (GRCm38.p 5). All differentially expressed mrnas and lncrnas were used for target identification. The regulatory relationships of miRNA-mRNA and miRNA-lncRNA were determined using RNA in situ hybridization techniques (setting the coordinates of the seed regions to 1 to 6, 2 to 7, 3 to 8, 4 to 9, and 5 to 10; mfe ≦ 25kcal/mol, p ≦ 0.01) and BLASTN (E value 1000, removing miRNA and miRNA in the forward alignment (alignment position greater than 6) and obtaining intersections.
Example three: construction of mouse model of schizophrenia:
the N-methyl-D-aspartate (NMDA) type glutamate receptor antagonist MK-801(Sigma-Aldrich, St. Louis, MO, USA) was dissolved in physiological saline and injected into mice by intraperitoneal injection for 7 days (0.6mg/kg, once a day). Mice in the same condition were injected with the same dose of physiological saline as a control. PET and qPCR experiments were performed within 1 week after injection.
Example four: positron Emission Tomography (PET):
according to normal operating procedures, mice were examined for PET imaging using the Siemens inventon PET/CT system (Siemens medical facility, knoxville, kentucky, usa) at the PET central washington hospital, university of compound denier. Mice were first anesthetized with isoflurane and the schizophrenic markers were injected via tail vein 50 minutes prior to detection18F-DTBZ into mice. Dynamic PET monitoring of mice was performed for 60 min.
Example five: brain imaging:
brain-related data was collected from the Shanghai Huashan Hospital for a panel of schizophrenic patients and healthy controls. All digital imaging and communications in medicine (DICOM) data were converted to NIfTI formatted files using DCM2NII (http:// scope. cas. sc. edu/rorden/mricron/index. html). The following two steps are used to establish the brain boundaries for analysis. First mapping the statistical parameters in MATLAB R2016a (SPM12, www.fil.ion.ucl.ac.uk/SPM); each image was normalized to a standard brain spatial map by the ICBM east asia template, and at the same time low frequency background noise was removed. Next, a FWHM value of 10X 10mm was applied3An isotropic gaussian smoothing kernel. T-test was performed using two sample data based on 18 schizophrenic patients and 26 healthy controls, comparing the groupsRegion of central interest (ROI). By setting the threshold to p<0.01 and applying a threshold of 20 pixels for error correction, the relevant area is located. Using Talairach Client, the individual and batch labels of the ROIs are found and created.
Example six: mouse brain RNA extraction and real-time quantitative PCR (qPCR)
Mice were quickly dissected in pre-cooled PBS and placed in RNA extraction solution after removal of the prefrontal lobe. RNA extraction was performed according to the protocol provided in the Promegas total RAN extraction kit. The total RNA extracted was then subjected to reverse transcription using a reverse transcription kit (DRR047) from TAKARA. Subsequently, the cDNA was used as a template, PCR primers for the target mRNA and a SYBR green Premix fluorescent quantitative PCR system from Yeasen were used to amplify the target mRNA on a stratagen MX3000P quantitative PCR instrument, and the Ct value of the sample was measured. And substituting the Ct value into a formula obtained by measuring a standard curve, and obtaining the content of the target mRNA in the sample by adopting an absolute quantitative calculation method. The miRNAqPCR assay results were normalized using GAPDH gene as an internal reference.
Example seven: blood sample RNA extraction and real-time quantitative PCR (qPCR)
Blood samples were collected using a blood collection tube without anticoagulant, and serum was taken after blood clotting for RNA extraction according to the protocol provided in the Yeasen total RAN extraction kit (Yeasen, shanghai, china). Then, the extracted total RNA was reverse transcribed using a reverse transcription kit from RiboBio, and then, the cDNA was used as a template, and PCR primers for the target mRNA and a qPCR SYBR Green Master Mix fluorescent quantitative PCR system from RiboBio were amplified on a stratagen MX3000P quantitative PCR instrument, and the Ct value of the sample was measured. And substituting the Ct value into a formula obtained by measuring a standard curve, and obtaining the content of the target mRNA in the sample by adopting an absolute quantitative calculation method. The results of the mRNAqPCR assay were normalized using the U6 gene as an internal reference.
The embodiment of the invention is helpful for people to find the molecular marker for the early diagnosis of schizophrenia by discussing the molecular mechanism of schizophrenia at the molecular level, and provides a new molecular target for the early prevention and treatment of schizophrenia. In the present invention, the whole genome difference of gene expression between schizophrenic patients and healthy control groups was first systematically analyzed using bioinformatics method, and 61 Differentially Expressed Genes (DEGs) were initially screened out among schizophrenic patients. Functional enrichment analysis of GO and KEGG shows that most genes are mainly enriched in synaptic and neurotransmitter conduction related functions. And then constructing a cerRNA network to analyze the interaction between the upstream key miRNA and the target gene thereof, wherein the research finds that the upstream miRNA number of the RASD2 gene is the maximum, and the central nervous system of the whole network is formed. The results show that different miRNAs regulate the expression of the DEGs, and miR-4763-3p can regulate the expression of a target gene RASD2, thereby influencing the occurrence and development of schizophrenia. Next, it was verified in mouse models and patients of schizophrenia, PET results revealed key brain regions of schizophrenia pathology such as prefrontal lobe, differential expression of RASD2 was confirmed in mouse models of schizophrenia through qPCR analysis, and key regulatory role of RASD2 in schizophrenia was demonstrated. Finally, qPCR analysis is carried out on blood samples of schizophrenic patients, and expression of RASD2 is reduced, and expression of miR-4763-3p expressed by target RASD2 is increased. In conclusion, the miR-4763-3p in the embodiment can be used as a potential biomarker and a therapeutic target of schizophrenia, and provides a theoretical basis for further clinical study of schizophrenia.
The embodiments of the present invention have been described above with reference to the accompanying drawings, but the present invention is not limited to the above embodiments, and various changes can be made according to the purpose of the invention, and any changes, modifications, substitutions, combinations or simplifications made according to the spirit and principle of the technical solution of the present invention should be replaced with equivalents as long as the object of the present invention is met, and the technical principle and the inventive concept of the present invention are not departed from the scope of the present invention.

Claims (7)

1. The application of miRNA molecular marker in schizophrenia detection is characterized in that: analyzing genome-wide differences in gene expression between schizophrenic patients and healthy controls by using bioinformatics methods, and then analyzing the signal pathways and functions in which these differential genes participate by functional enrichment of GO and KEGG; then establishing a cerRNA network to analyze the interaction between the upstream key miRNA and the target genes thereof, including the central target gene RASD2 and predict the optimal binding sites of the upstream key miRNA and the target genes; the expression of DEGs is regulated by miRNA, and hsa-miRNA-4763-3p targeting RASD2 gene is used as a potential biomarker and a therapeutic target of schizophrenia.
2. The use of the miRNA molecular marker of claim 1 in schizophrenia detection, wherein: functional enrichment analysis is carried out through gene ontology GO and gene and genome encyclopedia KEGG, and GO analysis comprises:
(1)17 genes were enriched in the biological process: including protein homooligomerization, modulation of potassium ion transport across membranes, axonal guidance, modulation of protein kinase activity, and retinal homeostasis;
(2)29 genes were enriched in cellular components: including cell junctions, cell surfaces, extracellular vesicles and golgi apparatus;
(3)13 genes are enriched in molecular functions: comprises voltage-gated potassium channel activity, polysaccharide binding, RNA polymerase II sequence specific DNA binding and potassium channel activity;
the KEGG analysis involved 10 DEGs participating in five important ways: graft versus host disease, allograft rejection, aldosterone-mediated sodium resorption, nicotine addiction, and type I diabetes.
3. The use of the miRNA molecular marker of claim 1 in schizophrenia detection, wherein: the expression of DEGs and related lncRNA is regulated by miRNA at the middle node of the cerRNA network, and the upstream miRNA of RASD2 gene comprises has-miRNA-4763-3p, hsa-miR-6820-5p, hsa-miR-6756-5p, hsa-miRNA-4695-5p and hss-miRNA-6769b-5p, so that the center of the whole network is formed.
4. The use of the miRNA molecular marker of claim 1 in schizophrenia detection, wherein: images were constructed and binding sites of target genes predicted using RNAhybrid and BLASTN.
5. The use of the miRNA molecular marker of claim 1 in schizophrenia detection, wherein: hsa-miRNA-4763-3p is predicted to be the optimal target site for binding to RASD 2.
6. The use of the miRNA molecular marker of claim 1 in schizophrenia detection, wherein: by injecting the 5-HT antagonist 18F-MPPF for specific detection of the expression profile of 5-HT receptors in schizophrenic patients, critical brain regions were analyzed.
7. The use of the miRNA molecular marker of claim 1 in schizophrenia detection, wherein: hsa-miRNA-4763-3p is used as a candidate biomarker for schizophrenia.
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何兴兰等: "miRNA-转录因子-靶基因前馈环路:肿瘤与动脉粥样硬化调控新机制", 生命的化学, vol. 33, no. 6, pages 621 - 626 *

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